DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure.
Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3.10(-5) for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.
Takai
T
T
Department of Biotechnology, Faculty of Engineering, Okayama University, Japan.
Ohmori
H
H
D000818:Animals; D002460:Cell Line; D003637:DEAE-Dextran; D004247:DNA / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D051379:Mice; D009997:Osmotic Pressure; D014162:Transfection
D004247:DNA; D003637:DEAE-Dextran